Analysis of nucleotide insertion opposite urea and translesion synthesis across urea by DNA polymerases

Taishu Kawada; Katsuhito Kino; Kyousuke Tokorodani; Ryuto Anabuki; Masayuki Morikawa; Takanobu Kobayashi; Kazuaki Ohara; Takayuki Ohshima; Hiroshi Miyazawa

doi:10.1186/s41021-022-00236-3

Analysis of nucleotide insertion opposite urea and translesion synthesis across urea by DNA polymerases

Genes Environ. 2022 Feb 15;44(1):7. doi: 10.1186/s41021-022-00236-3.

Authors

Taishu Kawada¹, Katsuhito Kino², Kyousuke Tokorodani¹, Ryuto Anabuki¹, Masayuki Morikawa¹, Takanobu Kobayashi¹, Kazuaki Ohara¹, Takayuki Ohshima³, Hiroshi Miyazawa¹

Affiliations

¹ Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University, 1314-1 Shido, Sanuki, Kagawa, 769-2193, Japan.
² Kagawa School of Pharmaceutical Sciences, Tokushima Bunri University, 1314-1 Shido, Sanuki, Kagawa, 769-2193, Japan. kkino@kph.bunri-u.ac.jp.
³ Faculty of Science and Engineering, Tokushima Bunri University, 1314-1 Shido, Sanuki, Kagawa, 769-2193, Japan.

Abstract

Urea (Ua) is produced in DNA as the result of oxidative damage to thymine and guanine. It was previously reported that Klenow fragment (Kf) exo^- incorporated dATP opposite Ua, and that DNA polymerase β was blocked by Ua. We report here the following nucleotide incorporations opposite Ua by various DNA polymerases: DNA polymerase α, dATP and dGTP (dATP > dGTP); DNA polymerase δ, dATP; DNA polymerase ζ, dATP; Kf exo^-, dATP; Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4), dGTP and dATP (dGTP > dATP); and DNA polymerase η, dCTP, dGTP, dATP, and dTTP (dCTP > dGTP > dATP > dTTP). DNA polymerases β and ε were blocked by Ua. Elongation by DNA polymerases δ and ζ stopped after inserting dATP opposite Ua. Importantly, the elongation efficiency to full-length beyond Ua using DNA polymerase η and Dpo4 were almost the same as that of natural DNA.

Keywords: Base pair; DNA polymerase; Elongation; Nucleotide incorporation; Oxidative DNA damage; Urea.

Grants and funding

17K00558/JSPS KAKENHI